CA2396210A1 - Method for cloning animals with targetted genetic alterations by transfer_of long-term cultured male or female somatic cell nuclei, comprising artificially-induced genetic alterations, to enucleated recipient cells - Google Patents
Method for cloning animals with targetted genetic alterations by transfer_of long-term cultured male or female somatic cell nuclei, comprising artificially-induced genetic alterations, to enucleated recipient cells Download PDFInfo
- Publication number
- CA2396210A1 CA2396210A1 CA002396210A CA2396210A CA2396210A1 CA 2396210 A1 CA2396210 A1 CA 2396210A1 CA 002396210 A CA002396210 A CA 002396210A CA 2396210 A CA2396210 A CA 2396210A CA 2396210 A1 CA2396210 A1 CA 2396210A1
- Authority
- CA
- Canada
- Prior art keywords
- cells
- cybrid
- cell
- fetus
- animal
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Abandoned
Links
- 210000004027 cell Anatomy 0.000 title claims abstract description 312
- 238000000034 method Methods 0.000 title claims abstract description 145
- 241001465754 Metazoa Species 0.000 title claims abstract description 118
- 210000001082 somatic cell Anatomy 0.000 title claims abstract description 81
- 238000010367 cloning Methods 0.000 title claims description 69
- 210000004940 nucleus Anatomy 0.000 title claims description 40
- 230000004077 genetic alteration Effects 0.000 title claims description 7
- 231100000118 genetic alteration Toxicity 0.000 title claims description 7
- 230000007774 longterm Effects 0.000 title abstract description 21
- 210000000287 oocyte Anatomy 0.000 claims abstract description 74
- 238000012546 transfer Methods 0.000 claims abstract description 69
- 210000003754 fetus Anatomy 0.000 claims description 69
- 210000001161 mammalian embryo Anatomy 0.000 claims description 39
- 238000012258 culturing Methods 0.000 claims description 23
- 230000004927 fusion Effects 0.000 claims description 23
- 210000001519 tissue Anatomy 0.000 claims description 23
- 241000124008 Mammalia Species 0.000 claims description 21
- 210000002459 blastocyst Anatomy 0.000 claims description 20
- 210000004291 uterus Anatomy 0.000 claims description 17
- 230000003213 activating effect Effects 0.000 claims description 14
- 238000004113 cell culture Methods 0.000 claims description 14
- 210000003855 cell nucleus Anatomy 0.000 claims description 14
- 108091093105 Nuclear DNA Proteins 0.000 claims description 13
- 241000894007 species Species 0.000 claims description 12
- 210000000056 organ Anatomy 0.000 claims description 9
- 238000000338 in vitro Methods 0.000 claims description 8
- 230000029803 blastocyst development Effects 0.000 claims description 7
- 238000002513 implantation Methods 0.000 claims description 7
- 230000008569 process Effects 0.000 claims description 7
- 230000037430 deletion Effects 0.000 claims description 4
- 238000012217 deletion Methods 0.000 claims description 4
- 230000006872 improvement Effects 0.000 claims description 4
- 229940043355 kinase inhibitor Drugs 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
- 150000007523 nucleic acids Chemical class 0.000 claims description 4
- 239000003909 protein kinase inhibitor Substances 0.000 claims description 4
- 238000012216 screening Methods 0.000 claims description 4
- 108091028043 Nucleic acid sequence Proteins 0.000 claims description 3
- 229940123573 Protein synthesis inhibitor Drugs 0.000 claims description 3
- 239000000007 protein synthesis inhibitor Substances 0.000 claims description 3
- 238000007792 addition Methods 0.000 claims description 2
- 238000001727 in vivo Methods 0.000 claims description 2
- 239000003112 inhibitor Substances 0.000 claims description 2
- 108020004707 nucleic acids Proteins 0.000 claims description 2
- 102000039446 nucleic acids Human genes 0.000 claims description 2
- 238000006467 substitution reaction Methods 0.000 claims description 2
- 210000002950 fibroblast Anatomy 0.000 description 36
- 210000002257 embryonic structure Anatomy 0.000 description 34
- 108020004414 DNA Proteins 0.000 description 21
- 230000022131 cell cycle Effects 0.000 description 21
- 239000002609 medium Substances 0.000 description 21
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 20
- 108090000623 proteins and genes Proteins 0.000 description 20
- 210000002966 serum Anatomy 0.000 description 20
- 230000035935 pregnancy Effects 0.000 description 18
- 230000004913 activation Effects 0.000 description 17
- 239000012091 fetal bovine serum Substances 0.000 description 17
- 230000007159 enucleation Effects 0.000 description 16
- 241000283690 Bos taurus Species 0.000 description 15
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 15
- 235000003642 hunger Nutrition 0.000 description 14
- 230000037351 starvation Effects 0.000 description 14
- 244000309466 calf Species 0.000 description 12
- 210000000349 chromosome Anatomy 0.000 description 12
- 230000000392 somatic effect Effects 0.000 description 12
- 244000309464 bull Species 0.000 description 11
- 238000004458 analytical method Methods 0.000 description 10
- 238000011161 development Methods 0.000 description 10
- 230000018109 developmental process Effects 0.000 description 10
- 230000031864 metaphase Effects 0.000 description 10
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 9
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 238000010790 dilution Methods 0.000 description 9
- 239000012895 dilution Substances 0.000 description 9
- 241000699666 Mus <mouse, genus> Species 0.000 description 8
- BVIAOQMSVZHOJM-UHFFFAOYSA-N N(6),N(6)-dimethyladenine Chemical compound CN(C)C1=NC=NC2=C1N=CN2 BVIAOQMSVZHOJM-UHFFFAOYSA-N 0.000 description 8
- 210000004748 cultured cell Anatomy 0.000 description 8
- YPHMISFOHDHNIV-FSZOTQKASA-N cycloheximide Chemical compound C1[C@@H](C)C[C@H](C)C(=O)[C@@H]1[C@H](O)CC1CC(=O)NC(=O)C1 YPHMISFOHDHNIV-FSZOTQKASA-N 0.000 description 8
- 230000013020 embryo development Effects 0.000 description 8
- 238000010353 genetic engineering Methods 0.000 description 8
- 102000011782 Keratins Human genes 0.000 description 7
- 108010076876 Keratins Proteins 0.000 description 7
- 108091092878 Microsatellite Proteins 0.000 description 7
- 230000001413 cellular effect Effects 0.000 description 7
- 210000001771 cumulus cell Anatomy 0.000 description 7
- 210000000805 cytoplasm Anatomy 0.000 description 7
- 238000010348 incorporation Methods 0.000 description 7
- 239000000243 solution Substances 0.000 description 7
- 238000010186 staining Methods 0.000 description 7
- 210000005048 vimentin Anatomy 0.000 description 7
- 241000283973 Oryctolagus cuniculus Species 0.000 description 6
- 210000002919 epithelial cell Anatomy 0.000 description 6
- 230000002068 genetic effect Effects 0.000 description 6
- 210000004602 germ cell Anatomy 0.000 description 6
- 230000037431 insertion Effects 0.000 description 6
- 238000003780 insertion Methods 0.000 description 6
- 239000000463 material Substances 0.000 description 6
- 238000002360 preparation method Methods 0.000 description 6
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 5
- 102000013127 Vimentin Human genes 0.000 description 5
- 108010065472 Vimentin Proteins 0.000 description 5
- 238000003556 assay Methods 0.000 description 5
- 230000002759 chromosomal effect Effects 0.000 description 5
- 230000000694 effects Effects 0.000 description 5
- 210000002308 embryonic cell Anatomy 0.000 description 5
- 210000001671 embryonic stem cell Anatomy 0.000 description 5
- 238000010449 nuclear transplantation Methods 0.000 description 5
- 210000004927 skin cell Anatomy 0.000 description 5
- PRDFBSVERLRRMY-UHFFFAOYSA-N 2'-(4-ethoxyphenyl)-5-(4-methylpiperazin-1-yl)-2,5'-bibenzimidazole Chemical compound C1=CC(OCC)=CC=C1C1=NC2=CC=C(C=3NC4=CC(=CC=C4N=3)N3CCN(C)CC3)C=C2N1 PRDFBSVERLRRMY-UHFFFAOYSA-N 0.000 description 4
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 102100027286 Fanconi anemia group C protein Human genes 0.000 description 4
- 230000010190 G1 phase Effects 0.000 description 4
- 102000004142 Trypsin Human genes 0.000 description 4
- 108090000631 Trypsin Proteins 0.000 description 4
- 238000001574 biopsy Methods 0.000 description 4
- 230000004663 cell proliferation Effects 0.000 description 4
- 238000012512 characterization method Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 4
- 239000012528 membrane Substances 0.000 description 4
- 238000000520 microinjection Methods 0.000 description 4
- 210000001672 ovary Anatomy 0.000 description 4
- 230000002035 prolonged effect Effects 0.000 description 4
- 230000008672 reprogramming Effects 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 230000001360 synchronised effect Effects 0.000 description 4
- 230000009261 transgenic effect Effects 0.000 description 4
- 239000012588 trypsin Substances 0.000 description 4
- 230000006820 DNA synthesis Effects 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 3
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 3
- GBOGMAARMMDZGR-UHFFFAOYSA-N UNPD149280 Natural products N1C(=O)C23OC(=O)C=CC(O)CCCC(C)CC=CC3C(O)C(=C)C(C)C2C1CC1=CC=CC=C1 GBOGMAARMMDZGR-UHFFFAOYSA-N 0.000 description 3
- 230000004075 alteration Effects 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 239000013553 cell monolayer Substances 0.000 description 3
- 239000006285 cell suspension Substances 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 238000012790 confirmation Methods 0.000 description 3
- GBOGMAARMMDZGR-JREHFAHYSA-N cytochalasin B Natural products C[C@H]1CCC[C@@H](O)C=CC(=O)O[C@@]23[C@H](C=CC1)[C@H](O)C(=C)[C@@H](C)[C@@H]2[C@H](Cc4ccccc4)NC3=O GBOGMAARMMDZGR-JREHFAHYSA-N 0.000 description 3
- GBOGMAARMMDZGR-TYHYBEHESA-N cytochalasin B Chemical compound C([C@H]1[C@@H]2[C@@H](C([C@@H](O)[C@@H]3/C=C/C[C@H](C)CCC[C@@H](O)/C=C/C(=O)O[C@@]23C(=O)N1)=C)C)C1=CC=CC=C1 GBOGMAARMMDZGR-TYHYBEHESA-N 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 239000012894 fetal calf serum Substances 0.000 description 3
- 238000000684 flow cytometry Methods 0.000 description 3
- 239000007850 fluorescent dye Substances 0.000 description 3
- 238000010363 gene targeting Methods 0.000 description 3
- 239000008103 glucose Substances 0.000 description 3
- 230000012010 growth Effects 0.000 description 3
- 238000003365 immunocytochemistry Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 230000004048 modification Effects 0.000 description 3
- 238000012986 modification Methods 0.000 description 3
- 210000003101 oviduct Anatomy 0.000 description 3
- 210000004508 polar body Anatomy 0.000 description 3
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 3
- 230000001850 reproductive effect Effects 0.000 description 3
- 230000007017 scission Effects 0.000 description 3
- 238000010374 somatic cell nuclear transfer Methods 0.000 description 3
- 238000002604 ultrasonography Methods 0.000 description 3
- BZTDTCNHAFUJOG-UHFFFAOYSA-N 6-carboxyfluorescein Chemical compound C12=CC=C(O)C=C2OC2=CC(O)=CC=C2C11OC(=O)C2=CC=C(C(=O)O)C=C21 BZTDTCNHAFUJOG-UHFFFAOYSA-N 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- 241000283725 Bos Species 0.000 description 2
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 description 2
- 102000018233 Fibroblast Growth Factor Human genes 0.000 description 2
- 108050007372 Fibroblast Growth Factor Proteins 0.000 description 2
- 206010064571 Gene mutation Diseases 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 241000283903 Ovis aries Species 0.000 description 2
- 108020002230 Pancreatic Ribonuclease Proteins 0.000 description 2
- 102000005891 Pancreatic ribonuclease Human genes 0.000 description 2
- 241001494479 Pecora Species 0.000 description 2
- 241001665167 Solter Species 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 210000004369 blood Anatomy 0.000 description 2
- 239000008280 blood Substances 0.000 description 2
- 238000010370 cell cloning Methods 0.000 description 2
- 239000002771 cell marker Substances 0.000 description 2
- 210000000170 cell membrane Anatomy 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 238000012411 cloning technique Methods 0.000 description 2
- 238000003501 co-culture Methods 0.000 description 2
- 210000002808 connective tissue Anatomy 0.000 description 2
- 238000010276 construction Methods 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- SDZRWUKZFQQKKV-JHADDHBZSA-N cytochalasin D Chemical compound C([C@H]1[C@@H]2[C@@H](C([C@@H](O)[C@H]\3[C@]2([C@@H](/C=C/[C@@](C)(O)C(=O)[C@@H](C)C/C=C/3)OC(C)=O)C(=O)N1)=C)C)C1=CC=CC=C1 SDZRWUKZFQQKKV-JHADDHBZSA-N 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 230000008175 fetal development Effects 0.000 description 2
- 230000001605 fetal effect Effects 0.000 description 2
- 229940126864 fibroblast growth factor Drugs 0.000 description 2
- 238000000799 fluorescence microscopy Methods 0.000 description 2
- 239000001963 growth medium Substances 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 238000012760 immunocytochemical staining Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000004962 mammalian cell Anatomy 0.000 description 2
- 239000003550 marker Substances 0.000 description 2
- 230000035800 maturation Effects 0.000 description 2
- 210000003632 microfilament Anatomy 0.000 description 2
- 230000011278 mitosis Effects 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 238000012758 nuclear staining Methods 0.000 description 2
- 230000016087 ovulation Effects 0.000 description 2
- 238000002559 palpation Methods 0.000 description 2
- 238000007747 plating Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 230000010076 replication Effects 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- HOZOZZFCZRXYEK-HNHWXVNLSA-M scopolamine butylbromide Chemical compound [Br-].C1([C@@H](CO)C(=O)OC2C[C@@H]3[N+]([C@H](C2)[C@@H]2[C@H]3O2)(C)CCCC)=CC=CC=C1 HOZOZZFCZRXYEK-HNHWXVNLSA-M 0.000 description 2
- 238000007390 skin biopsy Methods 0.000 description 2
- 210000001626 skin fibroblast Anatomy 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000001228 spectrum Methods 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- QTBSBXVTEAMEQO-UHFFFAOYSA-M Acetate Chemical compound CC([O-])=O QTBSBXVTEAMEQO-UHFFFAOYSA-M 0.000 description 1
- 241000251468 Actinopterygii Species 0.000 description 1
- 241001222053 Akabane virus Species 0.000 description 1
- 241000030939 Bubalus bubalis Species 0.000 description 1
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 description 1
- 241000282832 Camelidae Species 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 241001550206 Colla Species 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- 102000053602 DNA Human genes 0.000 description 1
- 230000010337 G2 phase Effects 0.000 description 1
- 108700012941 GNRH1 Proteins 0.000 description 1
- 230000010558 Gene Alterations Effects 0.000 description 1
- 229920002683 Glycosaminoglycan Polymers 0.000 description 1
- NMJREATYWWNIKX-UHFFFAOYSA-N GnRH Chemical class C1CCC(C(=O)NCC(N)=O)N1C(=O)C(CC(C)C)NC(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)CNC(=O)C(NC(=O)C(CO)NC(=O)C(CC=1C2=CC=CC=C2NC=1)NC(=O)C(CC=1NC=NC=1)NC(=O)C1NC(=O)CC1)CC1=CC=C(O)C=C1 NMJREATYWWNIKX-UHFFFAOYSA-N 0.000 description 1
- 239000000579 Gonadotropin-Releasing Hormone Substances 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 108010003272 Hyaluronate lyase Proteins 0.000 description 1
- 102000001974 Hyaluronidases Human genes 0.000 description 1
- 102100033421 Keratin, type I cytoskeletal 18 Human genes 0.000 description 1
- 108010066327 Keratin-18 Proteins 0.000 description 1
- 208000035752 Live birth Diseases 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 102000002151 Microfilament Proteins Human genes 0.000 description 1
- 108010040897 Microfilament Proteins Proteins 0.000 description 1
- 241000711408 Murine respirovirus Species 0.000 description 1
- 241000699670 Mus sp. Species 0.000 description 1
- 241001229135 Nassa Species 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 108020004511 Recombinant DNA Proteins 0.000 description 1
- 241000282849 Ruminantia Species 0.000 description 1
- 230000018199 S phase Effects 0.000 description 1
- 241000282887 Suidae Species 0.000 description 1
- HATRDXDCPOXQJX-UHFFFAOYSA-N Thapsigargin Natural products CCCCCCCC(=O)OC1C(OC(O)C(=C/C)C)C(=C2C3OC(=O)C(C)(O)C3(O)C(CC(C)(OC(=O)C)C12)OC(=O)CCC)C HATRDXDCPOXQJX-UHFFFAOYSA-N 0.000 description 1
- 108010077465 Tropocollagen Proteins 0.000 description 1
- 241000251539 Vertebrata <Metazoa> Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 210000001766 X chromosome Anatomy 0.000 description 1
- 210000002593 Y chromosome Anatomy 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 210000002718 aborted fetus Anatomy 0.000 description 1
- 206010000210 abortion Diseases 0.000 description 1
- 231100000176 abortion Toxicity 0.000 description 1
- NGCGMRBZPXEPOZ-HBBGHHHDSA-N acetic acid;(2s)-n-[(2s)-1-[[(2s)-1-[[(2s)-1-[[(2s)-1-[[2-[[(2s)-1-[[(2s)-1-[(2s)-2-[(2-amino-2-oxoethyl)carbamoyl]pyrrolidin-1-yl]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]amino]-2-oxoethyl]amino]-3-(4-hydroxyphenyl)- Chemical compound CC(O)=O.C([C@@H](C(=O)NCC(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCCN=C(N)N)C(=O)N1[C@@H](CCC1)C(=O)NCC(N)=O)NC(=O)[C@H](CO)NC(=O)[C@H](CC=1C2=CC=CC=C2NC=1)NC(=O)[C@H](CC=1NC=NC=1)NC(=O)[C@H]1NC(=O)CC1)C1=CC=C(O)C=C1 NGCGMRBZPXEPOZ-HBBGHHHDSA-N 0.000 description 1
- 239000002543 antimycotic Substances 0.000 description 1
- 229940045988 antineoplastic drug protein kinase inhibitors Drugs 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 210000004507 artificial chromosome Anatomy 0.000 description 1
- 238000011888 autopsy Methods 0.000 description 1
- 210000001109 blastomere Anatomy 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 150000001768 cations Chemical class 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 238000001516 cell proliferation assay Methods 0.000 description 1
- 108091092356 cellular DNA Proteins 0.000 description 1
- 210000003850 cellular structure Anatomy 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000001054 cortical effect Effects 0.000 description 1
- 238000005138 cryopreservation Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 229940002533 cystorelin Drugs 0.000 description 1
- 238000004163 cytometry Methods 0.000 description 1
- 210000004292 cytoskeleton Anatomy 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000034994 death Effects 0.000 description 1
- 231100000517 death Toxicity 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000017858 demethylation Effects 0.000 description 1
- 238000010520 demethylation reaction Methods 0.000 description 1
- 238000003745 diagnosis Methods 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 125000000118 dimethyl group Chemical group [H]C([H])([H])* 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 208000010515 dystocia Diseases 0.000 description 1
- 235000013601 eggs Nutrition 0.000 description 1
- 230000005611 electricity Effects 0.000 description 1
- 210000004920 epithelial cell of skin Anatomy 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 210000004996 female reproductive system Anatomy 0.000 description 1
- 230000004720 fertilization Effects 0.000 description 1
- 239000000834 fixative Substances 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000003209 gene knockout Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 238000005469 granulation Methods 0.000 description 1
- 230000003179 granulation Effects 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000003862 health status Effects 0.000 description 1
- 238000002744 homologous recombination Methods 0.000 description 1
- 230000006801 homologous recombination Effects 0.000 description 1
- 210000000003 hoof Anatomy 0.000 description 1
- 210000003917 human chromosome Anatomy 0.000 description 1
- 229960002773 hyaluronidase Drugs 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 238000010166 immunofluorescence Methods 0.000 description 1
- 238000005470 impregnation Methods 0.000 description 1
- 230000002779 inactivation Effects 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- PGHMRUGBZOYCAA-UHFFFAOYSA-N ionomycin Natural products O1C(CC(O)C(C)C(O)C(C)C=CCC(C)CC(C)C(O)=CC(=O)C(C)CC(C)CC(CCC(O)=O)C)CCC1(C)C1OC(C)(C(C)O)CC1 PGHMRUGBZOYCAA-UHFFFAOYSA-N 0.000 description 1
- PGHMRUGBZOYCAA-ADZNBVRBSA-N ionomycin Chemical compound O1[C@H](C[C@H](O)[C@H](C)[C@H](O)[C@H](C)/C=C/C[C@@H](C)C[C@@H](C)C(/O)=C/C(=O)[C@@H](C)C[C@@H](C)C[C@@H](CCC(O)=O)C)CC[C@@]1(C)[C@@H]1O[C@](C)([C@@H](C)O)CC1 PGHMRUGBZOYCAA-ADZNBVRBSA-N 0.000 description 1
- 239000002555 ionophore Substances 0.000 description 1
- 230000000236 ionophoric effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 239000010410 layer Substances 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 210000000265 leukocyte Anatomy 0.000 description 1
- 150000002632 lipids Chemical class 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 210000005075 mammary gland Anatomy 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 230000008774 maternal effect Effects 0.000 description 1
- 230000034217 membrane fusion Effects 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 238000000386 microscopy Methods 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 230000003547 miosis Effects 0.000 description 1
- 238000010369 molecular cloning Methods 0.000 description 1
- 238000012544 monitoring process Methods 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 230000001776 parthenogenetic effect Effects 0.000 description 1
- 230000032696 parturition Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 150000004633 phorbol derivatives Chemical class 0.000 description 1
- 239000002644 phorbol ester Substances 0.000 description 1
- XLCISDOVNFLSGO-VONOSFMSSA-N phorbol-12-myristate Chemical compound C([C@]1(O)C(=O)C(C)=C[C@H]1[C@@]1(O)[C@H](C)[C@H]2OC(=O)CCCCCCCCCCCCC)C(CO)=C[C@H]1[C@H]1[C@]2(O)C1(C)C XLCISDOVNFLSGO-VONOSFMSSA-N 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 239000003757 phosphotransferase inhibitor Substances 0.000 description 1
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 230000003389 potentiating effect Effects 0.000 description 1
- 239000002243 precursor Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 210000005000 reproductive tract Anatomy 0.000 description 1
- 230000001177 retroviral effect Effects 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 238000002741 site-directed mutagenesis Methods 0.000 description 1
- 238000010561 standard procedure Methods 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 150000003462 sulfoxides Chemical class 0.000 description 1
- 108091035539 telomere Proteins 0.000 description 1
- 102000055501 telomere Human genes 0.000 description 1
- 210000003411 telomere Anatomy 0.000 description 1
- -1 temperature change Substances 0.000 description 1
- IXFPJGBNCFXKPI-FSIHEZPISA-N thapsigargin Chemical compound CCCC(=O)O[C@H]1C[C@](C)(OC(C)=O)[C@H]2[C@H](OC(=O)CCCCCCC)[C@@H](OC(=O)C(\C)=C/C)C(C)=C2[C@@H]2OC(=O)[C@@](C)(O)[C@]21O IXFPJGBNCFXKPI-FSIHEZPISA-N 0.000 description 1
- 230000001052 transient effect Effects 0.000 description 1
- 229940045145 uridine Drugs 0.000 description 1
- 210000003462 vein Anatomy 0.000 description 1
- 230000000007 visual effect Effects 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- 238000003260 vortexing Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 230000003442 weekly effect Effects 0.000 description 1
- 210000004340 zona pellucida Anatomy 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N15/873—Techniques for producing new embryos, e.g. nuclear transfer, manipulation of totipotent cells or production of chimeric embryos
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K2217/00—Genetically modified animals
- A01K2217/05—Animals comprising random inserted nucleic acids (transgenic)
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Wood Science & Technology (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Environmental Sciences (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Plant Pathology (AREA)
- Developmental Biology & Embryology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Physics & Mathematics (AREA)
- Biophysics (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Molecular Biology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Applications Claiming Priority (5)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US17442400P | 2000-01-04 | 2000-01-04 | |
| US17438300P | 2000-01-04 | 2000-01-04 | |
| US60/174,383 | 2000-01-04 | ||
| US60/174,424 | 2000-01-04 | ||
| PCT/US2001/000394 WO2001049106A2 (en) | 2000-01-04 | 2001-01-04 | Method for cloning animals with targetted genetic alterations |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2396210A1 true CA2396210A1 (en) | 2001-07-12 |
Family
ID=26870158
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA002396210A Abandoned CA2396210A1 (en) | 2000-01-04 | 2001-01-04 | Method for cloning animals with targetted genetic alterations by transfer_of long-term cultured male or female somatic cell nuclei, comprising artificially-induced genetic alterations, to enucleated recipient cells |
Country Status (7)
| Country | Link |
|---|---|
| US (1) | US7071372B2 (zh) |
| EP (1) | EP1254210A4 (zh) |
| JP (1) | JP2003518936A (zh) |
| CN (1) | CN1293188C (zh) |
| AU (1) | AU2766401A (zh) |
| CA (1) | CA2396210A1 (zh) |
| WO (1) | WO2001049106A2 (zh) |
Families Citing this family (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US7253334B2 (en) * | 2000-12-22 | 2007-08-07 | Aurox, Llc | Methods for cloning non-human mammals using reprogrammed donor chromatin or donor cells |
| US20020142397A1 (en) | 2000-12-22 | 2002-10-03 | Philippe Collas | Methods for altering cell fate |
| WO2002062131A2 (en) * | 2001-02-02 | 2002-08-15 | Infigen, Inc. | Method of cloning transgenic mammalian animals using pseudonuclei |
| JP3842597B2 (ja) | 2001-08-14 | 2006-11-08 | 家畜受精卵移植技術研究組合 | Ｇ１期細胞の収集方法及び該細胞を用いた核移植方法 |
| US20030113910A1 (en) * | 2001-12-18 | 2003-06-19 | Mike Levanduski | Pluripotent stem cells derived without the use of embryos or fetal tissue |
| JP2006081542A (ja) * | 2004-08-18 | 2006-03-30 | Institute Of Physical & Chemical Research | クローン哺乳動物の作成方法 |
| WO2006018316A1 (en) * | 2004-08-20 | 2006-02-23 | Ingenium Pharmaceuticals Ag | Methods for the production of improved live stocks and disease models for therapeutic research |
| JP5327733B2 (ja) * | 2007-09-18 | 2013-10-30 | 国立大学法人名古屋大学 | 魚類胚の作製方法及びその利用 |
| EP2993234B1 (en) | 2013-04-30 | 2018-10-10 | Konkuk University Industrial Cooperation Corp. | Cmp-acetylneuraminic acid hydroxylase targeting vector, transgenic animal for xenotransplantation introduced with the vector, and method of manufacturing the same |
Family Cites Families (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5840493A (en) * | 1994-03-30 | 1998-11-24 | Mitokor | Mitochondrial DNA mutations that segregate with late onset diabetes mellitus |
| US5888498A (en) * | 1995-03-03 | 1999-03-30 | Mitokor | Cellular and animal models for diseases associated with mitochondrial defects |
| GB9517779D0 (en) | 1995-08-31 | 1995-11-01 | Roslin Inst Edinburgh | Biological manipulation |
| US5945577A (en) * | 1997-01-10 | 1999-08-31 | University Of Massachusetts As Represented By Its Amherst Campus | Cloning using donor nuclei from proliferating somatic cells |
| US6235969B1 (en) * | 1997-01-10 | 2001-05-22 | University Of Massachusetts | Cloning pigs using donor nuclei from non-quiescent differentiated cells |
| US6011197A (en) * | 1997-03-06 | 2000-01-04 | Infigen, Inc. | Method of cloning bovines using reprogrammed non-embryonic bovine cells |
| US6258998B1 (en) * | 1998-11-24 | 2001-07-10 | Infigen, Inc. | Method of cloning porcine animals |
| JP4373967B2 (ja) | 2005-07-08 | 2009-11-25 | 大王製紙株式会社 | マスク |
-
2001
- 2001-01-04 WO PCT/US2001/000394 patent/WO2001049106A2/en not_active Application Discontinuation
- 2001-01-04 CA CA002396210A patent/CA2396210A1/en not_active Abandoned
- 2001-01-04 US US09/755,204 patent/US7071372B2/en not_active Expired - Lifetime
- 2001-01-04 JP JP2001549484A patent/JP2003518936A/ja active Pending
- 2001-01-04 CN CNB018060552A patent/CN1293188C/zh not_active Expired - Lifetime
- 2001-01-04 AU AU27664/01A patent/AU2766401A/en not_active Abandoned
- 2001-01-04 EP EP01901802A patent/EP1254210A4/en not_active Withdrawn
Also Published As
| Publication number | Publication date |
|---|---|
| JP2003518936A (ja) | 2003-06-17 |
| EP1254210A2 (en) | 2002-11-06 |
| EP1254210A4 (en) | 2003-07-02 |
| CN1461339A (zh) | 2003-12-10 |
| WO2001049106A3 (en) | 2002-01-17 |
| WO2001049106A2 (en) | 2001-07-12 |
| US7071372B2 (en) | 2006-07-04 |
| AU2766401A (en) | 2001-07-16 |
| US20010037513A1 (en) | 2001-11-01 |
| CN1293188C (zh) | 2007-01-03 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Oback et al. | Cloning cattle | |
| Cibelli et al. | Trasgenic bovine chimeric offspring produced from somatic cell-derived stem-like cells | |
| US20040177390A1 (en) | Method of nuclear transfer | |
| Galli et al. | Mammalian leukocytes contain all the genetic information necessary for the development of a new individual | |
| WO1999005266A2 (en) | Trans-species nuclear transfer | |
| US6700037B2 (en) | Method of cloning porcine animals | |
| Lacham-Kaplan et al. | Fertilization of mouse oocytes using somatic cells as male germ cells | |
| JP4095898B2 (ja) | 人工染色体を含むトランスジェニック動物のクローニング | |
| US7071372B2 (en) | Method for cloning animals with targetted genetic alterations by transfer of long-term cultured male or female somatic cell nuclei, comprising artificially-induced genetic alterations, to enucleated recipient cells | |
| EP0774510A1 (en) | Ungulate eg cell | |
| AU2002252076A1 (en) | Cloning of transgenic animals comprising artificial chromosomes | |
| Eyestone et al. | Nuclear transfer from somatic cells: applications in farm animal species | |
| US20100293627A1 (en) | Method for cloning animals with targetted genetic alterations by transfer of long-term cultured male or female somatic cell nuclei, comprising artificially-induced genetic alterations, to enucleated recipient cells | |
| JP2006522609A (ja) | 動物の体細胞核移植と関係する紡錘体欠損を修正するための方法 | |
| US20040077077A1 (en) | Novel methods for the production of cloned mammals, mammals cloned according to the methods, and methods of use of same | |
| US20050149999A1 (en) | Methods for cloning mammals using remodeling factors | |
| AU2002248983B2 (en) | A method of nuclear transfer | |
| An et al. | Cloned calves produced by nuclear transfer from cultured cumulus cells | |
| JPH0614945A (ja) | 豚の繁殖方法 | |
| US20040064845A1 (en) | Method of cloning animals | |
| Heyman | Cloning and transgenesis in cattle: potential applications | |
| Gręda et al. | Developmental potential of selectively enucleated mouse zygotes reconstituted with embryonic cell, embryonic stem cell and somatic cell nuclei. | |
| AU2002248983A1 (en) | A method of nuclear transfer | |
| Spell et al. | Somatic cell cloning in the beef industry | |
| McLaughlin | Factors in the production of identical animals by nuclear transfer |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| FZDE | Discontinued |